High-Content Imaging

Molecular Devices ImageXpress Micro (replaced by IXM Confocal)

For high-content automated cellular imaging and analysis we are utilizing the Molecular Devices ImageXpress Micro (datasheet - for newer model IXMicroXL) with the live cell and phase contrast/brightfield option. We can image nearly any fluorophore with out collection of Semrock filters. See below for our filter configurations.

Please contact us for information on the IXMicro. For key card access to the IXMicro room (CCSR-0116) use iLabs.

iLabs IXMicro Schedule (click here to schedule training)
View only Calendar for non-registered users

ImageXpress Micro and Catalyst Express Robotic Arm

Features and Capabilities of the ImageXpress Micro

  • The IXMicro contains a fully automated inverted epifluorescence microscope -- Acquisition Guide -- User Guide
  •     Capable of scanning a 6-, 24-, 48-, 96- or 384-well microplates, slides or any other microplate sized container.
  •     Can recording multiple fluorescent images in a few minutes.
  • Can quantify morphological properties through the MetaXpress analysis software (Datasheet) -- Analysis Guide.
  • Our current ImageXpress Micro configuration permits the visualization of both fixed cells and live cells.
  •     The instrument has a temperature-, humidity-, and carbon dioxide-controlled chamber.
  • Can image non-fluorescent cells/objects with the phase contrast/brightfield option
  • Fully integrated to an external automated incubator (Thermo Cytomat 2C, 42 plates, Datasheet) to permit long-term or large-scale studies of cellular events via a robotic plate stacker (Thermo CataLyst Express Microplate HandlerDatasheet, User GuidePolara Scheduling Software, Polara User Guide) thereby increasing the throughput of this workstation.
  • Training provided by HTBC staff sign up here; iLabs IXMicro Schedule
  • Two MetaXpress analysis computers Analysis Computer1Analysis Computer2 also located in CCSR-0116
  • Prior to training, contact Jason Wu (x5-6004) to discuss what your imaging parameters. 
DataSheet  
Angiogenesis tube formation Cell health
Cell scoring Count nuclei
Granularity Live/dead
Mitotic Index Neurite outgrowth
Transfluor® Translocation — basic and enhanced

Preferred Imaging Plates

Wells

Vendor

Catalog Number

Objectives

24

Corning/Costar

 

All

96

Greiner

655090

All

96

Corning/Costar*

3904

All

384

Greiner

781091

All

384

Corning/Costar*

3712 or 4518 or 4581

All

 

*Secondary Preference

   

Technical Information

Objectives Excitation Filters Emission Filters Dichroics
4X 0.2 S Fluor 340_26nm CUBE 1-Quad 410/504/582/669 Quadband1
10X 0.3 Plan Fluor 427_10nm 440_40/521_21/607_34/700_45 410/504/582/669
20X 0.45 Plan Fluor ELWD 504_12nm CUBE 2-triple 464/542/639 Triband
40X 0.6 Plan Fluor ELWD 589_15nm 464_23/542_27/639_42 444/521/608
  387_11nm CUBE 3-CFP-LIVE  
Phase Objectives 485_20nm 483_32 458
4X PhL Plan Fluor DL 0.13 NA 560_25nm CUBE 4-YFP-LIVE  
10X Ph1 Plan Fluor DL 0.30 650_13nm 542-27 520
20X Ph1 Plan Fluor ELWD DM   CUBE 5-HcRed-LIVE  
    632-22 Triband
FIXED Cells Profile   CUBE 6-DAPI-FIXED (Pos. 3)  
    440-40 409
    CUBE 7-FITC-FIXED (Pos. 4)  
    525-30 506
    CUBE 8-TRITC-FIXED (Pos. 5)  
    607-36 Quadband
FRET Profile   CUBE 9-YFP-FRET (Pos. 4)  
(CFP-Live in Pos. 3)   542-27 458
    CUBE 10-Fura-2 (Pos. 5)  
    510-84nm Fura-2 (409)
Cy5 (Dedicated)   CUBE 11-Cy5 (Pos. 1)  
    692_40nm 660

Filter Sets

Label Name Excitation Emission Dichroic
Fura-2 340x 340_26nm D510-80nm/Fura-2 Fura-2
Fura-2 387x 387_11nm D510-80nm/Fura-2 Fura-2
CCF4 (FIXED) 387_11nm 525_30nm 506 (FITC-FIXED)
Ex396/Em580 387_11nm 607_36nm Quadband (TRITC-FIXED)
CFP 427_10nm 464_23/542_27/639_42 Triband
CFP-LIVE 427_10nm 483_32nm 458
FRET-LIVE 427_10nm 542_27nm 520 (YFP-LIVE)
YFP-FRET 427_10nm 542_27nm 458
YFP 504_12nm 464_23/542_27/639_42 Triband
YFP-LIVE 504_12nm 542_27nm 520
HcRED 589_15nm 464_23/542_27/639_42 Triband
HcRED-LIVE 589_15nm 632_22nm Triband
DAPI 387_11nm 440_40/521_21/607_34/700_45 Quadband
DAPI-FIXED 387_11nm 440_40nm 409
FITC 485_20nm 440_40/521_21/607_34/700_45 Quadband
FITC-FIXED 485_20nm 525_30nm 506
TRITC 560_25nm 440_40/521_21/607_34/700_45 Quadband
TRITC-FIXED 560_25nm 607_36nm Quadband
Ex488/Em617 485_20nm 607_36nm Quadband (TRITC-FIXED)
Ex510/Em580 485_20nm 607_36nm Quadband (TRITC-FIXED)
AlexaF594 (put HcRED_LIVE cube in Pos. 5) 560_25nm 632_22nm Triband
FRET YFP-ALX594 (put HcRED_LIVE cube in Pos. 5) 504_12nm 632_22nm Triband
Cy5 650_13nm 440_40/521_21/607_34/700_45 Quadband
Cy5 650_13nm 692_40nm 660

Optimized for these fluorophores

Cyan: CFP, AmCyan, SYTOX Blue, BOBO-1, BO-PRO-1
Yellow: YFP, Calcium Green-1, Fluo-3, Rhodamine 123
Red: HcRed, Cy3.5, Texas Red®, Alexa Fluor 594

Complete list of fluorophores for this filter set

Set description

This 5-filter "Pinkel" set is perfectly optimized for imaging CFP, YFP, and HcRed with high brightness, extremely low crosstalk, and superb signal-to-noise ratio. The set is also ideal for samples triple-labeled with CFP, YFP, and an orange or red dye, like Texas Red. It is comprised of three single-band exciters along with a triple-band beamsplitter and a triple-band emitter, and thus works best with an exciter filter wheel or slider.

Actual measured data from typical filters

Filter Average Transmission Center Wavelengths Bandwidths *
 Exciter 1 > 90% 427 nm 10 nm
 Exciter 2 > 90% 504 nm 12 nm
 Exciter 3 > 90% 589 nm 15 nm
 Emitter

> 90%, 90%, 90%

464, 542, 639 nm 23, 27, 42 nm
  Average Reflection Reflection Bands Transmission Bands
 Dichroic > 95%
> 95%
> 95%
420 - 430 nm
496 - 510 nm
579 - 596 nm
451 - 480 nm
530 - 561 nm
618 - 664 nm

 Exciter 1

 Dichroic

 Emitter

ASCII Data

ASCII Data

ASCII Data

 Exciter 2

 

 

ASCII Data

 

 

 Exciter 3

 

 

ASCII Data

Optimized for these fluorophores

Blue: DAPI, Hoechst, AMCA, Alexa Fluor 350
Green: FITC, GFP, rsGFP, BODIPY, Alexa Fluor 488        
Orange: TRITC, Cy3™, Texas Red®, MitoTracker Red, 
  Alexa Fluor 568 & 594
Red: Cy5™
, APC, TOTO-3, TO-PRO-3, Alexa Fluor 647 & 660

Complete list of fluorophores for this filter set

Set description

This 6-filter "Pinkel" quad-band set is designed for sequentially imaging DAPI, FITC, TRITC, and Cy5 (and other key fluorophores – see complete list above). This state-of-the-art set represents a break-through in quad-band performance, and has been tested by independent researchers to have up to four times the brightness, twice the signal-to-noise ratio and substantially lower crosstalk than any other such quad-band filter set on the market. The complete set is comprised of a quad-band beamsplitter and a quad-band emitter (these can be offered assembled in a microscope cube), plus four “no burn-out” single-band exciters that are intended to be mounted in an exciter filter wheel or slider.

Actual measured data from typical filters

Filter Average Transmission Center Wavelengths Bandwidths *
 Exciter 1 > 85% 387 nm 11 nm
 Exciter 2 > 90% 485 nm 20 nm
 Exciter 3 > 90% 560 nm 25 nm
 Exciter 4 > 90% 650 nm 13 nm
 Emitter

> 90%, 90%,
90%, 90%

440, 521,
607, 700 nm
40, 21,
34, 45 nm
  Average Reflection Reflection Bands Transmission Bands
 Dichroic > 95%
> 95%
> 95%
> 95%
381 - 392 nm
475 - 495 nm
547 - 572 nm
643 - 656 nm
420 - 460 nm
510 - 531 nm
589 - 623 nm
677 - 722 nm

 Exciter 1

 Dichroic

 Emitter

ASCII Data

ASCII Data

ASCII Data

 Exciter 2

 

 

ASCII Data

 

 

 Exciter 3

 

 

ASCII Data

 

 

 Exciter 4

 

 

ASCII Data